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Title: Proline: Mother Nature;s cryoprotectant applied to protein crystallography

Abstract

L-Proline is one of Mother Nature's cryoprotectants. Plants and yeast accumulate proline under freeze-induced stress and the use of proline in the cryopreservation of biological samples is well established. Here, it is shown that L-proline is also a useful cryoprotectant for protein crystallography. Proline was used to prepare crystals of lysozyme, xylose isomerase, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase for low-temperature data collection. The crystallization solutions in these test cases included the commonly used precipitants ammonium sulfate, sodium chloride and polyethylene glycol and spanned the pH range 4.6-8.5. Thus, proline is compatible with typical protein-crystallization formulations. The proline concentration needed for cryoprotection of these crystals is in the range 2.0-3.0 M. Complete data sets were collected from the proline-protected crystals. Proline performed as well as traditional cryoprotectants based on the diffraction resolution and data-quality statistics. The structures were refined to assess the binding of proline to these proteins. As observed with traditional cryoprotectants such as glycerol and ethylene glycol, the electron-density maps clearly showed the presence of proline molecules bound to the protein. In two cases, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase, proline binds in the active site. It is concluded that L-proline is an effective cryoprotectant for protein crystallography.

Authors:
; ; ; ; ;  [1]
  1. (UMC)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH)
OSTI Identifier:
1047307
Resource Type:
Journal Article
Journal Name:
Acta Crystallogr. D
Additional Journal Information:
Journal Volume: 68; Journal Issue: (8) ; 08, 2012
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; ACID PHOSPHATASE; AMMONIUM SULFATES; CRYSTALLIZATION; CRYSTALLOGRAPHY; CRYSTALS; DATA; DIFFRACTION; ELECTRON DENSITY; GLYCEROL; GLYCOLS; HISTIDINE; LYSOZYME; MOLECULES; OXIDOREDUCTASES; PLANTS; POLYETHYLENE GLYCOLS; PROLINE; PROTEINS; RANGE; RESOLUTION; SODIUM CHLORIDES; SOLUTIONS; STATISTICS; TEMPERATURE RANGE 0065-0273 K; XYLOSE; YEASTS

Citation Formats

Pemberton, Travis A., Still, Brady R., Christensen, Emily M., Singh, Harkewal, Srivastava, Dhiraj, and Tanner, John J. Proline: Mother Nature;s cryoprotectant applied to protein crystallography. United States: N. p., 2012. Web. doi:10.1107/S0907444912019580.
Pemberton, Travis A., Still, Brady R., Christensen, Emily M., Singh, Harkewal, Srivastava, Dhiraj, & Tanner, John J. Proline: Mother Nature;s cryoprotectant applied to protein crystallography. United States. doi:10.1107/S0907444912019580.
Pemberton, Travis A., Still, Brady R., Christensen, Emily M., Singh, Harkewal, Srivastava, Dhiraj, and Tanner, John J. Wed . "Proline: Mother Nature;s cryoprotectant applied to protein crystallography". United States. doi:10.1107/S0907444912019580.
@article{osti_1047307,
title = {Proline: Mother Nature;s cryoprotectant applied to protein crystallography},
author = {Pemberton, Travis A. and Still, Brady R. and Christensen, Emily M. and Singh, Harkewal and Srivastava, Dhiraj and Tanner, John J.},
abstractNote = {L-Proline is one of Mother Nature's cryoprotectants. Plants and yeast accumulate proline under freeze-induced stress and the use of proline in the cryopreservation of biological samples is well established. Here, it is shown that L-proline is also a useful cryoprotectant for protein crystallography. Proline was used to prepare crystals of lysozyme, xylose isomerase, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase for low-temperature data collection. The crystallization solutions in these test cases included the commonly used precipitants ammonium sulfate, sodium chloride and polyethylene glycol and spanned the pH range 4.6-8.5. Thus, proline is compatible with typical protein-crystallization formulations. The proline concentration needed for cryoprotection of these crystals is in the range 2.0-3.0 M. Complete data sets were collected from the proline-protected crystals. Proline performed as well as traditional cryoprotectants based on the diffraction resolution and data-quality statistics. The structures were refined to assess the binding of proline to these proteins. As observed with traditional cryoprotectants such as glycerol and ethylene glycol, the electron-density maps clearly showed the presence of proline molecules bound to the protein. In two cases, histidine acid phosphatase and 1-pyrroline-5-carboxylate dehydrogenase, proline binds in the active site. It is concluded that L-proline is an effective cryoprotectant for protein crystallography.},
doi = {10.1107/S0907444912019580},
journal = {Acta Crystallogr. D},
number = (8) ; 08, 2012,
volume = 68,
place = {United States},
year = {2012},
month = {9}
}